Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli NovaBlue (DE3) cells | Sorangium cellulosum |
Protein Variants | Comment | Organism |
---|---|---|
Q126E | the mutant shows 25.6% relative activity on maltose | Sorangium cellulosum |
Q126R | the mutant shows 11.4% relative activity on maltose | Sorangium cellulosum |
Q126S | the mutant shows 12.4% relative activity on maltose | Sorangium cellulosum |
Q219E/F220E | the mutant shows 5.2% relative activity on maltose | Sorangium cellulosum |
Q219K/F220C | the mutant shows 20% relative activity on maltose | Sorangium cellulosum |
Q219K/F220K | the mutant shows 29.3% relative activity on maltose | Sorangium cellulosum |
Q219N/F220K | the mutant shows 11.7% relative activity on maltose | Sorangium cellulosum |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.4 | - |
D-glucose | wild type enzyme, in 42 mM sodium phosphate buffer pH 7.5 at 37°C | Sorangium cellulosum | |
7.2 | - |
D-glucose | mutant enzyme Q219K/F220C, in 42 mM sodium phosphate buffer pH 7.5 at 37°C | Sorangium cellulosum | |
7.6 | - |
D-glucose | mutant enzyme Q219K/F220K, in 42 mM sodium phosphate buffer pH 7.5 at 37°C | Sorangium cellulosum | |
10.6 | - |
D-glucose | mutant enzyme Q219N/F220K, in 42 mM sodium phosphate buffer pH 7.5 at 37°C | Sorangium cellulosum | |
11 | - |
D-glucose | mutant enzyme Q126E, in 42 mM sodium phosphate buffer pH 7.5 at 37°C | Sorangium cellulosum | |
11.3 | - |
D-glucose | mutant enzyme Q219E/F220E, in 42 mM sodium phosphate buffer pH 7.5 at 37°C | Sorangium cellulosum | |
22.6 | - |
D-glucose | mutant enzyme Q126S, in 42 mM sodium phosphate buffer pH 7.5 at 37°C | Sorangium cellulosum | |
41.4 | - |
D-glucose | mutant enzyme Q126R, in 42 mM sodium phosphate buffer pH 7.5 at 37°C | Sorangium cellulosum |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | Ca2+ is needed for the correct folding and for the functionalization of pyrroloquinoline quinone as cofactor | Sorangium cellulosum |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
56000 | - |
calculated from amino acid sequence | Sorangium cellulosum |
63000 | - |
recombinant enzyme, SDS-PAGE | Sorangium cellulosum |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Sorangium cellulosum | - |
- |
- |
Sorangium cellulosum So ce56 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
HisTrap column chromatography, gel filtration | Sorangium cellulosum |
Renatured (Comment) | Organism |
---|---|
protein refolding is started on the column during elution in the presence of Ca2+. Refolding is finished by dialysis against 50 mM sodium potassium phosphate buffer pH 7.5 containing 0.003 mM CaCl2 and 0.002 mM pyrroloquinoline quinone. Refolding without Ca2+ and pyrroloquinoline quinone leads to an inactive enzyme | Sorangium cellulosum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-deoxy-D-glucose + 2,6-dichlorophenolindophenol | 3% activity compared to D-glucose | Sorangium cellulosum | 2-deoxy-D-glucono-1,5-lactone + reduced 2,6-dichlorophenolindophenol | - |
? | |
2-deoxy-D-glucose + 2,6-dichlorophenolindophenol | 3% activity compared to D-glucose | Sorangium cellulosum So ce56 | 2-deoxy-D-glucono-1,5-lactone + reduced 2,6-dichlorophenolindophenol | - |
? | |
D-allose + 2,6-dichlorophenolindophenol | 85% activity compared to D-glucose | Sorangium cellulosum | D-allono-1,5-lactone + reduced 2,6-dichlorophenolindophenol | - |
? | |
D-allose + 2,6-dichlorophenolindophenol | 85% activity compared to D-glucose | Sorangium cellulosum So ce56 | D-allono-1,5-lactone + reduced 2,6-dichlorophenolindophenol | - |
? | |
D-galactose + 2,6-dichlorophenolindophenol | 71% activity compared to D-glucose | Sorangium cellulosum | D-galactono-1,5-lactone + reduced 2,6-dichlorophenolindophenol | - |
? | |
D-galactose + 2,6-dichlorophenolindophenol | 71% activity compared to D-glucose | Sorangium cellulosum So ce56 | D-galactono-1,5-lactone + reduced 2,6-dichlorophenolindophenol | - |
? | |
D-glucose + 2,6-dichlorophenolindophenol | 100% activity | Sorangium cellulosum | D-glucono-1,5-lactone + reduced 2,6-dichlorophenolindophenol | - |
? | |
D-glucose + 2,6-dichlorophenolindophenol | 100% activity | Sorangium cellulosum So ce56 | D-glucono-1,5-lactone + reduced 2,6-dichlorophenolindophenol | - |
? | |
D-lactose + 2,6-dichlorophenolindophenol | 91% activity compared to D-glucose | Sorangium cellulosum | D-lactono-1,5-lactone + reduced 2,6-dichlorophenolindophenol | - |
? | |
D-lactose + 2,6-dichlorophenolindophenol | 91% activity compared to D-glucose | Sorangium cellulosum So ce56 | D-lactono-1,5-lactone + reduced 2,6-dichlorophenolindophenol | - |
? | |
D-maltose + 2,6-dichlorophenolindophenol | 71% activity compared to D-glucose | Sorangium cellulosum | D-maltono-1,5-lactone + reduced 2,6-dichlorophenolindophenol | - |
? | |
D-mannose + 2,6-dichlorophenolindophenol | 36% activity compared to D-glucose | Sorangium cellulosum | D-mannono-1,5-lactone + reduced 2,6-dichlorophenolindophenol | - |
? | |
D-xylose + 2,6-dichlorophenolindophenol | 86% activity compared to D-glucose | Sorangium cellulosum | D-xylono-1,5-lactone + reduced 2,6-dichlorophenolindophenol | - |
? | |
L-arabinose + 2,6-dichlorophenolindophenol | 97% activity compared to D-glucose | Sorangium cellulosum | L-arabinono-1,5-lactone + reduced 2,6-dichlorophenolindophenol | - |
? |
Synonyms | Comment | Organism |
---|---|---|
PQQ GDH | - |
Sorangium cellulosum |
pyrroloquinoline quinone dependent glucose dehydrogenase | - |
Sorangium cellulosum |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
- |
Sorangium cellulosum |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | 40 | at 25°C the enzyme activity is reduced to 50%, whereas at 40°C the activity is only reduced to 90% compared to the activity at 37°C | Sorangium cellulosum |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | 65 | 100% activity at 25-30°C, about 90% activity at 35°C, about 80% activity at 40-45°C, about 750% activity at 50°C, about 60% activity at 55°C, after incubation at 60°C for 1 h the enzyme still has 27% of its initial activity while loosing nearly all activity after treatment at 65°C | Sorangium cellulosum |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
121.5 | - |
D-glucose | mutant enzyme Q219K/F220C, in 42 mM sodium phosphate buffer pH 7.5 at 37°C | Sorangium cellulosum | |
197.1 | - |
D-glucose | mutant enzyme Q126E, in 42 mM sodium phosphate buffer pH 7.5 at 37°C | Sorangium cellulosum | |
227.2 | - |
D-glucose | mutant enzyme Q219N/F220K, in 42 mM sodium phosphate buffer pH 7.5 at 37°C | Sorangium cellulosum | |
305.9 | - |
D-glucose | mutant enzyme Q219E/F220E, in 42 mM sodium phosphate buffer pH 7.5 at 37°C | Sorangium cellulosum | |
341.6 | - |
D-glucose | mutant enzyme Q126R, in 42 mM sodium phosphate buffer pH 7.5 at 37°C | Sorangium cellulosum | |
445.1 | - |
D-glucose | mutant enzyme Q126S, in 42 mM sodium phosphate buffer pH 7.5 at 37°C | Sorangium cellulosum | |
1114 | - |
D-glucose | mutant enzyme Q219K/F220K, in 42 mM sodium phosphate buffer pH 7.5 at 37°C | Sorangium cellulosum | |
1779 | - |
D-glucose | wild type enzyme, in 42 mM sodium phosphate buffer pH 7.5 at 37°C | Sorangium cellulosum |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
the highest activity is observed with 42 mM sodium potassium phosphate buffer at pH 7.5 | Sorangium cellulosum |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
5.5 | 9 | - |
Sorangium cellulosum |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
pyrroloquinoline quinone | required for activity | Sorangium cellulosum |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
8.2 | - |
D-glucose | mutant enzyme Q126R, in 42 mM sodium phosphate buffer pH 7.5 at 37°C | Sorangium cellulosum | |
16.8 | - |
D-glucose | mutant enzyme Q219K/F220C, in 42 mM sodium phosphate buffer pH 7.5 at 37°C | Sorangium cellulosum | |
18 | - |
D-glucose | mutant enzyme Q126E, in 42 mM sodium phosphate buffer pH 7.5 at 37°C | Sorangium cellulosum | |
19.7 | - |
D-glucose | mutant enzyme Q126S, in 42 mM sodium phosphate buffer pH 7.5 at 37°C | Sorangium cellulosum | |
21.7 | - |
D-glucose | mutant enzyme Q219N/F220K, in 42 mM sodium phosphate buffer pH 7.5 at 37°C | Sorangium cellulosum | |
27 | - |
D-glucose | mutant enzyme Q219E/F220E, in 42 mM sodium phosphate buffer pH 7.5 at 37°C | Sorangium cellulosum | |
146.6 | - |
D-glucose | mutant enzyme Q219K/F220K, in 42 mM sodium phosphate buffer pH 7.5 at 37°C | Sorangium cellulosum | |
744 | - |
D-glucose | wild type enzyme, in 42 mM sodium phosphate buffer pH 7.5 at 37°C | Sorangium cellulosum |